WO2016050552A1

WO2016050552A1 - Process for producing a pufa-containing biomass which has high cell stability

Info

Publication number: WO2016050552A1
Application number: PCT/EP2015/071635
Authority: WO
Inventors: Horst Priefert; Jens Schneider; Christian Rabe; Amelia Claudia FIGUEIREDO-SILVA
Original assignee: Evonik Degussa Gmbh
Priority date: 2014-10-02
Filing date: 2015-09-22
Publication date: 2016-04-07
Also published as: CA2958466A1; CA2958466C; BR112017006838A2; DK180021B1; WO2016050965A1; CN107075540A; CL2017000754A1; ES2869980T3; EP3200605A1; US20170303561A1; EP3200605B1; ES2870093T3; CL2017000752A1; CA2958457C; EP3200602A1; US10842174B2; BR112017006838B1; DK201770206A1; DK201770208A1; DK180015B1

Abstract

According to the invention, when PUFA-producing cells are cultivated at high sulfate content, biomass that has a high cell stability and PUFAs that are effectively protected from oxidation can be obtained and can be processed further advantageously to obtain feedstuff.

Description

Process for the preparation of biomass containing PUFAs with high cell stability

The present invention relates to a method for producing a biomass containing PUFAs with high cell stability.

Processes for the production of polyunsaturated fatty acids (PUFAs) containing biomass are already described in the prior art. A problem with the biomass obtained is often the stability of the cell wall. The cell wall must not be too unstable, otherwise the oil contained could be released too early and the unsaturated fatty acids could be oxidized. On the other hand, the cell wall must not be too stable, otherwise the cells are not completely broken up in the manufacture of the feed and / or breaking the cells a very high energy input is required, so that the oil is not completely or only with very high energy input and so that high costs can be released from the cells.

It was therefore an object of the present invention to provide a process in which the biomass obtained has a cell stability which is suitable for the further processing of the biomass into the feed and which must not be too high or too low.

Surprisingly, it has been found according to the invention that the stability of the cell wall can be optimized by targeted addition of sulfate to the fermentation medium.

It has been found that by adding sulfate in an amount such that a sulfate concentration of 25 to 60 g / kg is established in the resulting biomass, optimal cell stability can be achieved.

In addition, it has been found that the biomass thus obtained can be processed at a very low energy input to a feed with high abrasion resistance and high water stability.

Furthermore, it has been found that the feed obtained using the biomass according to the invention can be used particularly advantageously for the cultivation of fish.

A further object of the present invention can therefore be seen in providing a biomass which, owing to its properties, is particularly suitable for further processing into a feedstuff. A first subject of the present invention is therefore a process for producing a polyunsaturated fatty acid (PUFAs) containing biomass

characterized in that for the production of the biomass, a cultivation of microorganisms is carried out in a fermentation medium containing sulfate in an amount such that in the resulting biomass, a sulfate concentration, based on the dry mass, of 25 to 60 g / kg. The sulfate concentration in the resulting biomass is preferably 25 to 50 g / kg, in particular 25 to 40 g / kg, particularly preferably 25 to 35 g / kg, in each case based on the dry mass.

Another object of the present invention is also a PUFAs containing biomass, which is obtainable by a method according to the invention.

The present invention also relates to a biomass containing PUFAs, which has a sulfate content of 25 to 60 g / kg, based on the dry matter, and is preferably obtainable by a previously described process. The sulfate content is preferably 25 to 50 g / kg, in particular 25 to 40 g / kg, particularly preferably 25 to 35 g / kg, in each case based on the dry mass.

According to the invention, "sulfate content" is to be understood as meaning the total content of sulfate, ie the content of free and bound, in particular organically bound, sulfate It is to be assumed that the majority of the sulfate contained in the biomass is known as

Component of exopolysaccharides is involved, which are involved in the formation of the cell wall of the microorganisms.

The determination of the sulfate content is carried out according to the invention preferably by determining the sulfur content of the resulting biomass, since the majority of the sulfur contained in the biomass is due to the sulfate contained. Sulfur due to other sources is negligible due to the amount of sulphate it contains. From the amount of sulfur detected can thus be determined easily the amount of sulfate contained.

The sulfur content of the biomass is determined here preferably by elemental analysis according to DIN EN ISO 1 1885. For the analysis of the sulfur content of the biomass, appropriate aliquots of the sample are preferably digested with nitric acid and hydrogen peroxide at 240 ° C prior to analysis to ensure the free accessibility of the sulfur contained therein. Another object of the present invention is therefore also a method for

Production of a biomass containing polyunsaturated fatty acids (PUFAs), characterized in that for the production of the biomass a cultivation of microorganisms takes place in a fermentation medium which contains sulfate in an amount, so that in the resulting biomass by elemental analysis according to DIN EN ISO 1 1885 a Sulfur content of 8 to 20 g / kg, based on the dry mass, can be detected. The sulfur content in the resulting biomass is in this case preferably 8 to 17 g / kg, in particular 8 to 14 g / kg, particularly preferably 8 to 12 g / kg, in each case based on the dry matter. A further subject of the present invention is therefore also a PUFA-containing biomass, characterized in that elemental analysis according to DIN EN ISO 1885 shows a sulfur content of 8 to 20 g / kg, based on the dry mass, can be detected. The sulfur content in the resulting biomass is in this case preferably 8 to 17 g / kg, in particular 8 to 14 g / kg, particularly preferably 8 to 12 g / kg, in each case based on the dry mass.

According to the invention, the phosphorus content of biomasses according to the invention is preferably 1 to 6 g / kg, in particular 2 to 5 g / kg, relative to the dry matter. The phosphorus content is preferably also determined by elemental analysis according to DIN EN ISO 1 1885. The biomass according to the invention preferably comprises cells, and preferably consists essentially of those cells which already naturally produce PUFAs, but it may also be cells which are produced by appropriate genetic engineering

Were enabled to produce PUFAs. The production can be autotrophic, mixotrophic or heterotrophic. The cells of the biomass are preferably those which produce PUFAs heterotrophically. According to the invention, the cells are preferably algae, fungi, in particular yeasts, or protists. Particularly preferred are cells of microbial algae or fungi.

Yeasts of Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon and Lipomyces are particularly suitable as cells of oil-producing yeasts. A biomass according to the invention preferably comprises cells, and preferably consists essentially of such cells, the taxon Labyrinthulomycetes (labyrinthulea, slime fungi, mucous membranes), in particular those of the family Thraustochytriaceae. The family of Thraustochytriaceae includes the genera Althomia, Aplanochytrium, Elnia, Japonochytrium, Schizochytrium, Thraustochytrium, Aurantiochytrium,

Oblongichytrium and ulkenia. Particularly preferred are cells of the genera Thraustochytrium, Schizochytrium, Aurantiochytrium or Oblongichytrium, especially those of the genus Aurantiochytrium. A particularly preferred strain is the strain Aurantiochytrium limacinum SR21 (IFO 32693). The biomass according to the invention is preferably the product of a fermentative cultivation process. In addition to the cells to be disrupted, the biomass may also contain constituents of the fermentation medium. These ingredients may be, in particular, salts, antifoaming agents and unreacted carbon source and / or nitrogen source. The cell content in this biomass is preferably at least 70% by weight, preferably at least 75% by weight. The cell content in the biomass may, if appropriate, be carried out before the

Cell disruption process by appropriate washing steps, for example, be increased to at least 80 or at least 90 wt .-%. However, the obtained biomass can also be used directly in the cell disruption process. The cells of the biomass are preferably characterized in that they have a PUFA content of at least 20% by weight, preferably at least 30% by weight, in particular at least 35% by weight, based in each case on the cell dry mass.

In a preferred embodiment, a large part of the lipids is present in the form of triglycerides, preferably at least 50% by weight, in particular at least 75% by weight and in a particularly preferred embodiment at least 90% by weight of the lipids contained in the cell Form of triglycerides are present.

Preferably, at least 10% by weight, in particular at least 20% by weight, particularly preferably 20 to 60% by weight, in particular 20 to 40% by weight, of the fatty acids present in the cell are PUFAs. According to the invention, polyunsaturated fatty acids (PUFAs) are fatty acids which have at least two CC double bonds. Among the PUFAs, according to the invention, highly unsaturated fatty acids (HUFAs) are preferred. Be under HUFAs understood according to the invention fatty acids having at least four CC double bonds.

The PUFAs can be present in the cell in free form or in bound form. Examples of the presence in bound form are phospholipids and esters of PUFAs,

in particular monoacyl, diacyl and triacylglycerides. In a preferred

Embodiment, a majority of PUFAs is in the form of triglycerides, preferably at least 50 wt .-%, in particular at least 75 wt .-% and in a particularly preferred embodiment, at least 90 wt .-% of the PUFAs contained in the cell in the form of triglycerides available. Preferred PUFAs are omega-3 fatty acids and omega-6 fatty acids, omega-3 fatty acids being particularly preferred. Preferred omega-3 fatty acids hereby comprise eicosapentaenoic acid (EPA, 20: 5ω-3), in particular the (5Z, 8Z, 11Z, 14Z, 17Z) -ecoso-5,8,11,14,17-pentaenoic acid , and the docosahexaenoic acid (DHA, 22: 6ω-3), in particular the (4Z, 7Z, 10Z, 13Z, 16Z, 19Z) docosa-4,7, 10,13,16, 19-hexaenoic acid, wherein the

Docosahexaenoic acid is particularly preferred.

Methods of making the PUFA-containing cells, in particular of the order Thraustochytriales, have been extensively described in the art (see, e.g.

WO91 / 07498, WO94 / 08467, WO97 / 37032, WO97 / 36996, WO01 / 54510). The preparation is usually carried out by culturing cells in the presence of a carbon source and a nitrogen source in a fermenter. It can biomass densities of more than 100 grams per liter and production rates of more than 0.5 grams of lipid per liter per hour can be achieved. The process is preferably carried out as a so-called fed-batch process, i. that the carbon and nitrogen rates are incrementally supplied during the fermentation. The lipid production can be induced after reaching the desired biomass by different measures,

for example, by limiting the nitrogen source, the carbon source or the oxygen content, or combinations thereof.

The source of carbon is both alcoholic and non-alcoholic

Carbon sources into consideration. Examples of alcoholic carbon sources are methanol, ethanol and isopropanol. Examples of non-alcoholic carbon sources are fructose, glucose, sucrose, molasses, starch and corn syrup. Suitable sources of nitrogen include both inorganic and organic nitrogen sources. Examples of inorganic nitrogen sources are nitrates and ammonium salts, especially ammonium sulfate and ammonium hydroxide. Examples of organic

Nitrogen sources are amino acids, especially glutamate, and urea. The achievement of the desired sulfate content in the resulting biomass can be carried out according to the invention in different ways.

For example, in a so-called batch process, the required amount of sulfate can already be completely submitted at the beginning. The amount of sulfate needed is easily calculated as the cells used to form the biomass almost completely assimilate the sulfate.

When using a so-called fed-batch process, alternatively, the amount of sulfate required in the course of the fermentation can be added or submitted according to a part of the sulfate and the remainder be metered in the course of the fermentation.

In particular, if it turns out in the course of fermentation that the amount of produced biomass exceeds the originally calculated value, can

subsequent metered addition of sulfate to ensure that the resulting biomass has sufficient cell stability.

The sulfate salt used according to the invention preferably sodium sulfate, ammonium sulfate or magnesium sulfate and mixtures thereof. The chloride content is during the fermentation, with respect to the liquid

Fermentation medium including the biomass contained, preferably always below 3 g / kg, in particular below 1 g / kg, more preferably below 400 mg / kg

Fermentation medium.

In addition to sulfates and optionally used chlorides, other salts, in particular selected from sodium carbonate, sodium bicarbonate, soda ash or inorganic phosphorus compounds, may optionally also be used in the fermentation.

If further salts are used, they are preferably used in an amount such that each individual during the fermentation, with respect to the liquid fermentation medium including the biomass contained, in each case in an amount of always present less than 10 g / kg, in particular less than 5 g / kg, more preferably less than 3 g / kg in the fermentation medium.

According to the invention, the total salt content in the fermentation medium, including the biomass contained, is always below 35 g / kg, in particular below 30 g / kg, during the entire fermentation. With particular preference, the total salt content during the entire fermentation, with respect to the liquid fermentation medium including the biomass contained, is between 10 and 35 g / kg, in particular between 12 and 30 g / kg.

The sulphate content in the fermentation medium including the biomass contained according to the invention is preferably always between 5 and 16 g / kg in the course of the entire fermentation.

In addition, in the fermentation medium and organic phosphorus compounds and / or known growth stimulating substances, such as yeast extract or

Corn steep liquor, are added to positively influence the fermentation. The fermentation of the cells is preferably carried out at a pH of 3 to 1 1, in particular 4 to 10, and preferably at a temperature of at least 20 ° C, in particular 20 to 40 ° C, particularly preferably at least 30 ° C. A typical one

Fermentation process takes up to about 100 hours.

The cells are preferably fermented according to the invention up to a biomass density of at least 50, 60 or 70 g / l, in particular at least 80 or 90 g / l, particularly preferably at least 100 g / l. The data refer here to the content of dry biomass with respect to the total volume of the fermentation broth after completion of the fermentation. The content of bio-dry matter is determined by filtering off the biomass from the fermentation broth, then washing with water, then complete drying - for example in the microwave - and finally determination of the dry weight.

Preferably, after the harvest of the cells, or possibly even shortly before the harvest of the cells, the cells are pasteurized in order to kill the cells and inactivate enzymes which could promote the breakdown of the lipids. After completion of the fermentation, the biomass is harvested. By centrifugation, filtration, decantation or solvent evaporation, much of the Fermentation medium can be separated from the biomass. The

Solvent evaporation is preferably carried out using a drum dryer, a tunnel dryer, spray drying or vacuum evaporation. The

Solvent evaporation can in particular also by using a

Rotary evaporator, a thin film evaporator or a falling film evaporator done. As an alternative to solvent evaporation, for example, the

Reverse osmosis for the concentration of the fermentation broth into consideration. Subsequently, the resulting biomass is optionally further dried, preferably by

Fluid bed granulation. By drying, the moisture content is preferably reduced to below 15% by weight, in particular below 10% by weight, particularly preferably below 5% by weight.

According to the invention, "dry mass" is preferably to be understood as meaning a biomass which has a moisture content of less than 10% by weight, in particular less than 5% by weight a nozzle spray drying method, as described for example in EP13176661.0.

If necessary, silica may be added to the biomass during drying as an anti-caking agent in order to convert it into a more manageable state. The biomass-containing fermentation broth and the silica are for this purpose

preferably sprayed in the respective drying zone. Alternatively, the biomass is preferably mixed with the silica only after drying. Reference is also made in this regard to the patent application EP13187631 .0.

In a preferred embodiment, a biomass to be used according to the invention after drying has a concentration of silica, in particular hydrophilic or hydrophobic silica, of 0.2 to 10% by weight, in particular 0.5 to 5% by weight, especially 0.5 up to 2% by weight.

By drying, a free-flowing, finely divided or coarse-grained product, preferably granules, is preferably obtained. Optionally, a product having the desired grain size can be obtained from the granules obtained by sieving or dust separation. If a free-flowing finely divided powder has been obtained, this can optionally be converted by suitable compacting or granulation processes into a coarse-grained, free-flowing, storable and substantially dust-free product.

In this subsequent granulation or compaction may optionally conventional organic or inorganic adjuvants or carriers such as starch, gelatin, cellulose derivatives or similar substances commonly used in the food or

Feed processing can be used as binding, gelling or thickening agent used.

"Free-flowing" according to the invention is to be understood as meaning a powder which can flow out of a series of glass outlet vessels with differently sized outlet openings at least from the vessel with the opening 5 mm (Klein: Soaps, Oils, Fats, Wachse 94, 12 (1968)) ,

According to the invention, "finely divided" is to be understood as meaning a powder having a predominant fraction (> 50%) with a particle size of 20 to 100 micrometers in diameter. "Coarse-grained" means according to the invention a powder having a predominant fraction (> 50%) of a particle size of 100 to 2500 microns in diameter.

"Dust-free" is to be understood according to the invention as a powder which contains only small amounts (<10%, preferably <5%) of particle sizes below 100 micrometers.

The determination of the grain or particle sizes is carried out according to the invention preferably by methods of laser diffraction spectrometry. The methods to be used are described in the textbook "Particle Size Measurement in Laboratory Practice" by R. H. Müller and R. Schuhmann, Wissenschaftliche Verlagsgesellschaft Stuttgart (1996) and in the textbook "Introduction to Particle Technology" by M. Rhodes, Verlag Wiley & Sons (1998). If different methods are applicable, preference is given to the first applicable methodology from the textbook of R.H. Müller and R. Schuhmann zur

Particle size measurement applied.

The products obtained by drying processes according to the invention have

preferably a fraction of at least 80% by weight, in particular at least 90% by weight, particularly preferably at least 95% by weight, of particles having a particle size of 100 to 3500 micrometers, preferably 100 to 3000 micrometers, especially 100 to 2500 micrometers. The products of a fluidized-bed granulation process obtained according to the invention preferably have a proportion of at least 80% by weight, in particular at least 90% by weight, particularly preferably at least 95% by weight, of particles having a particle size of 200 to 3500 micrometers, preferably 300 to 3000 microns, especially 500 to 2500 microns.

By contrast, the products of a spray-drying process obtained according to the invention preferably have a proportion of at least 80% by weight, in particular at least 90% by weight, particularly preferably at least 95% by weight, of particles having a particle size of from 100 to 500 micrometers, preferably from 100 to 400 microns, especially 100 to 300 microns.

The products obtained according to the invention of a spray-drying process and subsequent granulation process preferably have a proportion of at least 80 wt .-%, in particular at least 90 wt .-%, particularly preferably at least 95 wt .-%, of particles having a particle size of 100 to 1000 micrometers. The proportion of dust, i. Particles having a particle size of less than 100 micrometers, is preferably at most 10 wt .-%, in particular at most 8 wt .-%, particularly preferably at most 5 wt .-%, especially at most 3 wt .-%.

The bulk density of the products according to the invention is preferably 400 to 800 kg / m ³ , more preferably 450 to 700 kg / m ³ . According to the invention, it has further been found that a biomass according to the invention, with a low energy input, becomes a high-quality feed

Oil loading capacity, high abrasion resistance and high water stability can be further processed.

A further subject of the present invention is therefore also a feed comprising a biomass according to the invention as well as other feed ingredients.

The other feed ingredients are hereby preferably selected from

proteinaceous, carbohydrate-containing, nucleic acid-containing and lipid-soluble components and optionally further fatty-containing components and furthermore from other additives such as minerals, vitamins, pigments and amino acids. Besides nutrient substances, structuring substances may also be present in order to improve the texture or the appearance of the feedstuff. Furthermore you can For example, binders are used to influence the consistency of the feed. A preferred component that is both a nutrient and a structurant is starch.

According to the invention, a feed according to the invention or a composition used for producing a feed according to the invention is preferably characterized in that it comprises a biomass according to the invention in an amount of 2 to 24% by weight, preferably 4 to 22% by weight, in particular 9 to 20 Wt .-%, especially 1 1 to 18 wt .-%, contains.

This feed or the composition used to prepare the feed preferably further comprises at least one, preferably all, of the following

Properties on: a) a total protein content of 33 to 67 wt .-%, preferably 39 to 61 wt .-%, in particular 44 to 55 wt .-%; b) a total fat content of 5 to 25% by weight, preferably 8 to 22% by weight,

in particular 10 to 20 wt .-%, especially 12 to 18 wt .-%; c) a total starch content of at most 25% by weight, in particular not more than 20

Wt .-%, preferably 6 to 17 wt .-%, particularly preferably 8 to 14 wt .-%; d) a content of polyunsaturated fatty acids (PUFAs) of 2 to 13% by weight,

preferably 3 to 1 1 wt .-%, in particular 4 to 10 wt .-%, especially 5.5 to 9 wt .-%; e) a content of omega-3 fatty acids of 1 to 7 wt .-%, preferably 1, 5 to 5.5 wt .-%, in particular 2 to 5 wt .-%, especially 2.5 to 4.5 wt .-%; f) a content of DHA of 0.5 to 3 wt .-%, preferably 0.8 to 2.8 wt .-%,

in particular 1 to 2.8 wt .-%, especially 1, 3 to 2.4 wt .-%, in particular 1, 3 to 2.2 wt .-%.

A preferred article according to the invention is therefore also a feed or a composition suitable for producing the feed, which has at least one, preferably all, of the following properties: a) a total protein content of 33 to 67% by weight, preferably 39 to 61 wt .-%, in particular 44 to 55 wt .-%; b) a total fat content of 5 to 25 wt .-%, preferably 8 to 22 wt .-%, in particular 10 to 20 wt .-%, especially 12 to 18 wt .-%; c) a total starch content of at most 25% by weight, in particular not more than 20

A preferred article according to the invention is therefore also a feed or a composition suitable for producing the feed, which has at least one, preferably all, of the following properties: a) a total protein content of 33 to 67% by weight, preferably 39 to 61 wt .-%, in particular 44 to 55 wt .-%, b) a total fat content of 5 to 25 wt .-%, preferably 8 to 22 wt .-%, in particular 10 to 20 wt .-%, especially From 12 to 18% by weight; c) a total starch content of at most 25% by weight, in particular not more than 20

Wt .-%, preferably 6 to 17 wt .-%, particularly preferably 8 to 14 wt .-%; d) a content of biomass according to the invention, in particular one

Labyrinthulea biomass according to the invention, preferably a Thraustochytriaceae biomass according to the invention, of 2 to 24% by weight, preferably 4 to 22

Wt .-%, in particular 9 to 20 wt .-%, especially 1 1 to 18 wt .-%; e) a content of polyunsaturated fatty acids (PUFAs) of 2 to 13% by weight,

preferably 3 to 1 1 wt .-%, in particular 4 to 10 wt .-%, especially 5.5 to 9 wt .-%; f) a content of omega-3 fatty acids of 1 to 7 wt .-%, preferably 1, 5 to 5.5 wt .-%, in particular 2 to 5 wt .-%, especially 2.5 to 4.5 wt .-%; g) a content of DHA of 0.5 to 3 wt .-%, preferably 0.8 to 2.8 wt .-%,

A preferred article according to the invention is therefore also a feed or a composition suitable for producing the feed which has at least one, preferably all, of the following properties: a) a total protein content of from 33 to 67% by weight, preferably 39 to 61% by weight, especially 40 to 50% by weight; b) a total fat content of 5 to 25 wt .-%, preferably 8 to 22 wt .-%, in particular 10 to 20 wt .-%, especially 12 to 18 wt .-%; c) a total starch content of at most 25% by weight, in particular not more than 20

Wt .-%, preferably 6 to 17 wt .-%, particularly preferably 8 to 14 wt .-%; d) a content of an inventive Aurantiochytrien- biomass, preferably an Aurantiochytrium limacinum biomass according to the invention, especially an Aurantiochytrium limacinum SR21 biomass according to the invention, from 2 to 24% by weight, preferably 4 to 22 wt .-%, in particular 9 to 20 wt .-%, especially 1 1 to 18 wt .-%; e) a content of polyunsaturated fatty acids (PUFAs) of 2 to 13% by weight,

By extrusion of the aforementioned compositions, an extrudate having an abrasion resistance of at least 91%, in particular at least 92, 93 or 94% can be obtained. These extrudates are a preferred subject of the present invention. The abrasion resistance was determined according to the invention in the following manner: The dried extrudate (4 mm in diameter and 4 mm in length) was subjected to mechanical stress using the Holmen pellet tester NHP100 (Borregaard Lignotech, Hüll, UK). Prior to performing the test, the samples were sieved to

possibly to remove adhering fine particles. The prepared samples (100 g) were then introduced into the pellet tester using a 2.5 mm filter screen. The pellets were then transported at high air velocity (about 70 mbar) for 30 seconds through a tube with a right-angled tube bend. The

Test parameters are specified by the device. Subsequently, the abrasion was determined by weighing. Abrasion resistance was reported as PDI (Pellet Durability Index), defined as the amount of sample left in the filter screen after the test was run. The test was carried out in each case with three samples and then the mean value was formed.

It has proved to be particularly advantageous according to the invention if the extrusion takes place at an energy input of 12-28 Wh / kg, in particular 14-26 Wh / kg, particularly preferably 16-24 Wh / kg, especially 18-22 Wh / kg ,

In the extrusion process is preferably a screw or

Twin-screw extruder used. The extrusion process is preferably carried out at a temperature of 80-220 ° C., in particular 80-130 ° C., especially 95-1 10 ° C., a pressure of 10-40 bar, and a wave circulation speed of 100-1,000 rpm, in particular 300 ° C. 700 rpm. The residence time of the introduced mixture is preferably 5 to 30 seconds, in particular 10 to 20 seconds.

The extrusion process may optionally comprise a compaction and / or a compression step. Before carrying out the extrusion process, the components are preferably intimately mixed. This is preferably done in a drum with

Shovels is equipped. In this mixing step takes place in a preferred

Embodiment, a water vapor injection, in particular to the swelling of

preferably contained starch effect. The steam injection is in this case preferably carried out at a pressure of 1 to 5 bar, more preferably at a pressure of 2 to 4 bar. The other food or feed ingredients are preferably comminuted, if necessary, before mixing with the algal biomass to ensure that a homogeneous mixture is obtained in the mixing step. The comminution of the other food or feed ingredients can be done, for example, using a hammer mill.

The extrudate produced preferably has a diameter of 1 to 14 mm, preferably 2 to 12 mm, in particular 2 to 6 mm, and preferably also has a length of 1 to 14 mm, preferably 2 to 12 mm, in particular 2 to 6 mm , The length of the extrudate is adjusted by using a cutting tool during extrusion. The length of the extrudate is preferably selected to be approximately equal to the diameter of the extrudate. The diameter of the extrudate is determined by selecting the sieve diameter.

In a preferred embodiment according to the invention is followed by the

Extrusion process, the loading of the resulting extrudate with oil. For this purpose, the extrudate is preferably first dried to a moisture content of at most 5 wt .-%. The loading of the extrusion product with oil can according to the invention

For example, by inserting the extrudate in oil or spraying the extrudate with oil done, it is according to the invention, however, preferably by vacuum coating.

In this way, feeds are obtained, the biomasses according to the invention

preferably in an amount of 2 to 22 wt .-%, in particular 4 to 20 wt .-%, particularly preferably 8 to 18 wt .-%, especially 10 to 16 wt .-%, included.

Accordingly, these feedstocks preferably furthermore have at least one, preferably all, of the following properties: a) a total protein content of from 30 to 60% by weight, preferably from 35 to 55% by weight, in particular from 40 to 50% by weight; b) a total fat content of 15 to 35 wt .-%, preferably 18 to 32 wt .-%, in particular 20 to 30 wt .-%, especially 22 to 28 wt .-%; c) a total starch content of at most 25% by weight, in particular not more than 20

Wt .-%, preferably 5 to 15 wt .-%, particularly preferably 7 to 13 wt .-%; a content of polyunsaturated fatty acids (PUFAs) of 2 to 12 wt .-%, preferably 3 to 10 wt .-%, in particular 4 to 9 wt .-%, especially 5 to 8 wt .-%; a content of omega-3 fatty acids of 1 to 6 wt .-%, preferably 1, 5 to 5 wt .-%, in particular 2 to 4.5 wt .-%, especially 2.5 to 4 wt .-% ; a content of DHA of 0.5 to 3 wt .-%, preferably 0.8 to 2.5 wt .-%, in particular 1 to 2.5 wt .-%, especially 1, 2 to 2.2 wt. -%, In particular 1, 2 to 2.0 wt .-%.

A preferred article according to the invention is therefore also a feed, in particular an extrudate, which has at least one, preferably all, of the following properties: a) a total protein content of 30 to 60% by weight, preferably 35 to 55% by weight , in particular 40 to 50 wt .-%; b) a total fat content of 15 to 35 wt .-%, preferably 18 to 32 wt .-%, in particular 20 to 30 wt .-%, especially 22 to 28 wt .-%; c) a total starch content of at most 25% by weight, in particular not more than 20

Wt .-%, preferably 5 to 15 wt .-%, particularly preferably 7 to 13 wt .-%; d) a content of polyunsaturated fatty acids (PUFAs) of 2 to 12% by weight,

preferably 3 to 10 wt .-%, in particular 4 to 9 wt .-%, especially 5 to 8 wt .-%; e) a content of omega-3 fatty acids of 1 to 6 wt .-%, preferably 1, 5 to 5 wt .-%, in particular 2 to 4.5 wt .-%, especially 2.5 to 4 wt. -%; f) a content of DHA of 0.5 to 3 wt .-%, preferably 0.8 to 2.5 wt .-%,

in particular 1 to 2.5 wt .-%, especially 1, 2 to 2.2 wt .-%, in particular 1, 2 to 2.0 wt .-%.

A preferred article according to the invention is therefore also a feed, in particular an extrudate, which has at least one, preferably all, of the following properties: a) a total protein content of 30 to 60 wt .-%, preferably 35 to 55 wt .-%, in particular 40 to 50 wt .-%; b) a total fat content of 15 to 35 wt .-%, preferably 18 to 32 wt .-%, in particular 20 to 30 wt .-%, especially 22 to 28 wt .-%; c) a total starch content of at most 25% by weight, in particular not more than 20

Wt .-%, preferably 5 to 15 wt .-%, particularly preferably 7 to 13 wt .-%; d) a content of a biomass according to the invention, in particular a

Labyrinthulea biomass according to the invention, preferably one

Thraustochytriaceae biomass of the invention, from 2 to 22 wt .-%, preferably 4 to 20 wt .-%, in particular 8 to 18 wt .-%, especially 10 to 16

Wt .-%; e) a content of polyunsaturated fatty acids (PUFAs) of 2 to 12% by weight,

preferably 3 to 10 wt .-%, in particular 4 to 9 wt .-%, especially 5 to 8 wt .-%; f) a content of omega-3 fatty acids of 1 to 6 wt .-%, preferably 1, 5 to 5

Wt .-%, in particular 2 to 4.5 wt .-%, especially 2.5 to 4 wt .-%; g) a content of DHA of 0.5 to 3 wt .-%, preferably 0.8 to 2.5 wt .-%,

in particular 1 to 2.5 wt .-%, especially 1, 2 to 2.2 wt .-%, in particular 1, 2 to 2.0 wt .-%. A preferred article according to the invention is therefore also a feed, in particular an extrudate, which has at least one, preferably all, of the following properties: a) a total protein content of 30 to 60 wt.%, Preferably 35 to 55 wt. -%, in particular 40 to 50 wt .-%; b) a total fat content of 15 to 35 wt .-%, preferably 18 to 32 wt .-%, in particular 20 to 30 wt .-%, especially 22 to 28 wt .-%; c) a total starch content of at most 25% by weight, in particular not more than 20

Wt .-%, preferably 5 to 15 wt .-%, particularly preferably 7 to 13 wt .-%; a content of an inventive Aurantiochytrien biomass, preferably an inventive Aurantiochytrium limacinum biomass, especially an Aurantiochytrium limacinum SR21 biomass according to the invention, from 2 to 22

Wt .-%, preferably 4 to 20 wt .-%, in particular 8 to 18 wt .-%, especially 10 to 16 wt .-%; d) a content of polyunsaturated fatty acids (PUFAs) of 2 to 12% by weight,

preferably 3 to 10 wt .-%, in particular 4 to 9 wt .-%, especially 5 to 8 wt .-%; e) a content of omega-3 fatty acids of 1 to 6 wt .-%, preferably 1, 5 to 5 wt .-%, in particular 2 to 4.5 wt .-%, especially 2.5 to 4 wt. -%; f) a content of DHA of 0.5 to 3 wt .-%, preferably 0.8 to 2.5 wt .-%, in particular 1 to 2.5 wt .-%, especially 1, 2 to 2.2 Wt .-%, in particular 1, 2 to 2.0 wt .-%.

The aforementioned extrudates obtainable by oil coating preferably have a water stability of at least 96%, in particular at least 97 or 98%, and constitute a further preferred subject of the present invention.

The determination of water stability was carried out essentially as described by Baeverfjord et al. (2006; Aquaculture 261, 1335-1345), with minor

Modifications. 10 g samples of the extrudate (4 mm in length and 4 mm in diameter) were placed in 6.5 mm diameter, 0.3 mm mesh metallic infusion baskets (Inox, Germany). The infusion cups were then placed in a plastic tub with water so that the samples were completely covered with water. The pan was then subjected to a 30 minute shaking agitation for 30 minutes using the multi-orbital shaker PSU-20I (Biosan, Latvia). Thereafter, the samples were carefully dried with blotting paper and then weighed before and after being subjected to oven drying at a temperature of 105 ° C for 24 hours. Water stability was calculated as the difference in dry weight of the sample before and after incubation in water and as a percentage of the dry weight of the sample used prior to incubation with water.

As a fat-containing component, according to the invention, in addition to the biomass to be used according to the invention, fats, in particular oils, animal as well as plant-based Origin are used. Vegetable oils according to the invention are, in particular, suitable as a fatty component, for example soybean oil, rapeseed oil, sunflower seed oil, flaxseedol or palm oil and mixtures thereof. In addition, if appropriate, fish oil can also be used as a fat-containing component in small amounts.

Preferably, a feed according to the invention which has an abrasion resistance of at least 96, 97 or 98%, vegetable oils in an amount of 3 to 18 wt .-%, in particular 5 to 15 wt .-%, especially 7 to 13 parts by weight. %. As described above, the vegetable oil here is preferably subsequently applied to the extrudate, in particular by vacuum coating.

For example, soy protein, pea protein, wheat gluten or corn gluten and mixtures thereof can be used as the proteinaceous component.

As a protein-containing component, which additionally contains fats, can be used, for example, fish meal, krill flour, shellfish, squid or Schrimpsschalen. These are summarized below under the term "marine flour." In a preferred embodiment, a feed according to the invention comprises marine flour, preferably fishmeal, in an amount of from 3 to 18% by weight, in particular from 5 to 15% by weight, especially 7 to 13 wt .-%.

As carbohydrate-containing component, for example, wheat flour,

Sunflower meal or soybean meal, as well as mixtures thereof.

When using feed according to the invention, in particular an oil-coated extrudate according to the invention, in animal breeding, it has been found that this promotes the growth of the animals to a particular extent and improves the stress state of the animals. A further subject of the present invention is also a method of raising animals, characterized in that a feed according to the invention is administered thereto.

The present invention is in particular a method for

Increasing the growth of animals, characterized in that a feed according to the invention is administered to them. Another object of the present invention is in particular also a method for increasing the proportion of omega-3 fatty acids, in particular DHA, in the muscle tissue of animals, characterized in that a feed according to the invention is administered to them. Preferably, in the method according to the invention, the feed is administered at least every two days, preferably at least once a day.

Another object of the present invention is also the use of a feed according to the invention for increasing the growth in animals.

Another object of the present invention is also the use of a feed according to the invention for increasing the proportion of omega-3 fatty acids in muscle tissue in animals.

Another object of the present invention is also the use of a feed according to the invention for improving the health of animals, in particular for improving the stress state of animals. Another object of the present invention is also the use of a feed according to the invention to enable stress-reduced breeding of the animals.

The cultured animals which are fed with a feed according to the invention are preferably poultry, pigs or cattle. However, the cultured animals are particularly preferably marine animals, particularly preferably fin fish or crustaceans. These include in particular carp, tilapia, catfish, tuna, salmon, trout, baramundi, bream, perch, cod, shrimp, lobster, crab, shrimp and crayfish. Particularly preferably, the cultured animals are salmon. The most popular salmon species are Atlantic salmon, red salmon, masu salmon, king salmon, keta salmon, silver salmon, Danube salmon, pacific salmon and pink salmon.

The cultured animals may in particular also be fish which are subsequently processed into fishmeal or fish oil. The fish are preferably herring, pollack, menhaden, anchovies, capelin or cod. The fish meal or fish oil thus obtained can in turn be used in aquaculture for the breeding of food fish or crustaceans. However, the cultured animals may also be microorganisms used as feed in aquaculture. These microorganisms may be, for example, nematodes, crustaceans or rotifers.

The cultivation of the marine animals can take place in ponds, tanks, basins or even in delimited areas in the sea or in lakes, in particular in cages or net pens. Cultivation can be used to grow the finished fish, but can also be used to raise juveniles, which are then released to replenish the stock of wild fish.

In salmon farming, it is preferable first to cultivate herbs to fresh salmon in freshwater tanks or artificial streams of water and then to continue breeding in the sea in floating cages or net pens, which are preferably anchored in bays or fjords.

The feed according to the invention is accordingly preferably a feed for use in the cultivation of the aforementioned animals.

embodiments

Example 1: Production of the biomass by fermentation of Aurantiochytrium limacinum SR21 in media with different sodium sulfate content

The cultivation of the cells was carried out for about 75 h in a feed process using a steel fermenter with a fermenter volume of 2 liters at a total

Starting mass of 712 g and a total final mass of 1, 3 - 1, 5 kg. During the process, a glucose solution (570 g / kg glucose) was added ("fed-batch process").

The composition of the starting media was as follows: medium 1: 20 g / kg glucose; 4 g / kg yeast extract; 2 g / kg ammonium sulfate; 2.46 g / kg

Magnesium sulfate (heptahydrate); 0.45 g / kg potassium chloride; 4.5 g / kg

potassium; 0.1 g / kg thiamine (HCl); 5 g / kg trace element solution.

Medium2: Same as medium 1 plus 8 g / kg sodium sulfate

Medium 3: Same as medium 1 plus 12 g / kg sodium sulfate Medium4: Same as medium 1 plus 16 g / kg sodium sulfate

The composition of the trace element solution was as follows: 35 g / kg hydrochloric acid (37%); 1.86 g / kg manganese chloride (tetrahydrate); 1, 82 g / kg zinc sulfate (heptahydrate); 0.818 g / kg sodium EDTA; 0.29 g / kg boric acid; 0.24 g / kg of sodium molybdate (dihydrate); 4.58 g / kg calcium chloride (dihydrate); 17.33 g / kg iron sulfate (heptahydrate); 0.15 g / kg copper chloride (dihydrate).

The cultivation was carried out under the following conditions: culturing temperature 28 ° C; Aeration rate 0.5 vvm, stirrer speed 600 - 1950 rpm, control of the pH in the growth phase at 4.5 with the aid of ammonia water (25% v / v). The following biomass densities were obtained: 100 g / l (medium 1), 1 l 1 g / l (medium 2), 1 14 g / l (medium 3), 1 16 g / l (medium 4).

After cultivation, the fermentation broths were heated for 20 minutes at 60 ° C to prevent further activity of the cells.

Example 2: Determination of the DHA Content of the Biomass

After inactivation, the resulting biomasses were subjected to fatty acid analysis. For this purpose, in each case 0.2 to 0.5 ml of fermentation broth were provided with 1 ml of internal standard and made up with 9 ml of a methanol / chloroform solution (1: 2, v / v). The samples were treated for 10 minutes in an ultrasonic bath. The samples were then evaporated to dryness at 50 ° C. in the thermoblock under nitrogen. To the

Dry residues were each added 2 ml of 0.5N KOH and incubated for 15 min at 100 ° C. The samples were then cooled to room temperature, mixed with 2 ml of 0.7N HCl and 1 ml of boron trifluoride solution (14% BF 3 in methanol) and incubated at 100 ° C. for a further 15 min. After cooling to room temperature, the samples were each extracted with a mixture of 3 ml of water and 2 ml of heptane. After centrifugation for 1 min at 2000 rpm, 1 ml each of the upper phase was transferred to a GC vial and analyzed by gas chromatography.

The analysis showed that all four biomasses contained a DHA content of more than 32% by weight relative to the total amount of fatty acids contained. Example 3: Drying of the biomass obtained

The cooled biomass-containing fermentation broths with different Na ₂ SO ₄ contents according to Example 1 were each separately subjected to spray drying.

Spray drying was carried out using a Buchi Mini Spray Dryer B-290 (diameter of nozzle tip: 0.7 mm, flow of spray air: 742 L / h, flow rate of aspirator: 35 m ³ / h, temperature of inlet air: 220 ° C, temperature of outlet air: 80 ° C).

Example 4: Determination of Sulfate and DHA Levels of Spray Dried Samples

The samples obtained by spray-drying the fermentation broths with different Na 2 SO 4 contents were subjected to a sulfate or sulfur determination and a DHA concentration determination. The DHA determination was carried out as described under Example 2. The sulfur content was determined in accordance with DIN EN ISO 1 1885.

Table 1: Analysis of spray-dried samples

Example 4: Determination of Clumping Tendency

The fermentation broths obtained after fermentation in media with different sodium sulphate contents showed marked differences in the spray-drying process and the resulting spray-dried material had different degrees of agglomeration, which was based on released oil. The biomass produced by fermentation in the

Media 3 and 4 were obtained, showed a significantly lower tendency to clump than the biomass obtained by fermentation in media 1 and 2. This is evidence of the increased cell stability of the cells in the biomass in question.

Example 5: Drying of the sulfate-rich biomass from Example 1 for the purpose

Feed production The biomass of Example 1 obtained in the sulfate-rich medium 4 was subjected to two-stage drying to produce feed: First, the fermentation broth was concentrated by evaporation to a dry matter content of about 20% by weight. Subsequently, spray drying of the concentrated fermentation broth was carried out using a Production Minor ™ Spray Dryer (GEA NIRO) at one

Inlet temperature of the drying air of 340 ° C. By spray-drying, a powder having a dry matter content of more than 95% by weight was thus obtained.

Example 6: Production of a feed by extrusion

The feed mixtures shown in Table 3 were prepared. In addition to the biomass according to the invention to be used according to Example 5, two other commercially available Labyrinthulea biomasses and fish oil were tested as the currently still common source of omega-3 fatty acids for comparison.

The preparation of the feed mixtures was carried out in each case by mixing the

Components - except oils - using a double helix mixer (Model 500L, TGC Extrusion, France). The mixtures thus obtained were then comminuted using a hammer mill (model SH1, Hosokawa-Alpine, Germany) to particle sizes below 250 μηη.

Table 2: Feed compositions used in the extrusion (data in% by weight)

For the extrusion each 140 kg per feed were used. The extrusion was carried out by means of a twin-screw extruder (CLEXTRAL BC45) with a

Screw diameter of 55.5 mm and a maximum flow rate of 90-100 kg / h performed. Pellets with a size of 4.0 mm (diameter and length) extruded. For this purpose, the extruder was equipped with a high-speed cutter in order to convert the product into the intended pellet size.

Various extrusion parameters have now been tested to find out under which extrusion conditions an optimum oil loading capacity of the resulting extrudate can be obtained. It has surprisingly been found that an optimal oil loading capacity can be achieved with a very low energy input. The energy input was significantly lower than with the use of fish oil. Furthermore, in the case of an algae biomass which is preferably to be used according to the invention, the optimum energy contribution was again significantly lower than for commercially available algae biomasses. The results are shown in Table 3.

Table 3: Energy inputs for producing pellets with the desired

Oil loading capacity

The size "SME" is the specific mechanical energy calculated as follows:

_TT . Test SS

U x 1 x cos

SME (Wh / kg) = Max SS

qs With

U: working voltage of the motor (in the present case 460 V) I: current of the motor (A) cos Φ: theoretical power of the extruder motor (in this case 0.95) Test SS: test speed (rpm) of the rotating screws

Max SS: maximum speed (267 rpm) of the rotating screw Qs: inlet flow rate of the feed pulp (kg / h)

After extrusion, the extrudate was dried in a vibrating fluid bed dryer (Model DR100, TGC Extrusion, France). Finally, after the extrudate had cooled, an oil coating was applied by vacuum coating (vacuum coater PG-10VCLAB, Dinnisen, The Netherlands).

Example 7 Determination of Abrasion Resistance and Water Stability of the Feedstuff of Example 6

Abrasion resistance was evaluated as follows: The dried extrusion was subjected to mechanical stress prior to loading with oil using the Holmen Pellet Tester (Borregaard Lignotech, Hüll, UK). Prior to performing the test, the samples were screened to remove any adhering fines. The prepared samples (100 g) were then introduced into the pellet tester using a 2.5 mm filter screen. The pellets were then treated with high

Air speed for 30 seconds through a tube, the right-angled pipe bend had. Subsequently, the abrasion was determined by weighing. The

Abrasion resistance was reported as PDI (Pellet Durability Index), defined as the amount of sample remaining on the filter screen in percent. The test was carried out in each case with three samples and then the mean value was formed.

Water stability was performed on the ol loaded samples. The method was essentially carried out as described by Baeverfjord et al. (2006, Aquaculture 261, 1335-1345), with minor modifications. 10 g samples were placed in metal infusion baskets with a mesh size of 0.3 mm. The infusion baskets were

then put in a plastic tub with water, so that the samples completely with Water were covered. The pan was then subjected to a shaking agitation of 30 shaking units per minute for 30 minutes. Thereafter, the samples were carefully dried with blotting paper and then weighed before and after being subjected to oven drying at a temperature of 105 ° C for 24 hours. Water stability was calculated as the difference in dry weight of the sample before and after incubation in water and as a percentage of the dry weight of the sample used prior to incubation with water.

The results are shown in the following Table 4.

It can be seen that a feed according to the invention which contains a biomass according to the invention has a significantly higher abrasion resistance and water stability than feed containing a commercially available labyrinthulea biomass or fish oil as the source of omega-3 fatty acids.

Example 8: Production of feed for feeding experiments

In each case feeds were prepared by extrusion of the biomass from Example 5, containing, based on the dry matter, 42.5 wt .-% total protein and 24 wt .-% total lipid and have a pellet size of 3 mm.

A total of three different feed formulations were prepared (diet 1, 2 and 3). The control formulation "Diet 1" contained 1 1, 0 wt .-% fish oil In the recipe "diet 2" the fish oil was partially replaced (about 50%) by Aurantiochytrien biomass, by 9.1 wt .-% Biomass were added and for the amount of fish oil was reduced to 5.5 wt .-%. In the recipe "diet 3", the fish oil was completely replaced by aurantiochrytric biomass by adding 16 wt.% Biomass and at the same time increasing the amount Rapeseed oil was increased from 8.2 to 9.9 wt .-%. Differences in total weight were offset by the amount of added wheat.

The individual components of the feed are shown in the following table.

Table 5: Recipes used in cultivation

The individual components were intimately mixed with each other except for the oils and then extruded using a twin-screw extruder (Wenger TX 52, Wenger, USA) using a 2 mm diameter die. The extrudates were about 1 hour in a carousel dryer (Paul Klöckner, Process Engineering GmbH, Germany) at 65 ° C to a water content of 7 dried to 8 wt .-%. Subsequently, the extrudates were dried overnight at room temperature before the oils were applied by vacuum coating (Dinnissen, Sevenum, The Netherlands).

Example 9 Feeding experiments with the formulations from Example 8

In order to perform the feeding experiments, three containers each containing a total of 83.6 g of young salmon with a total weight of salmon of 4 kg per container were fed with each of these formulations for a total of 12 weeks.

The total weight of salmon per container increased during this period from 4 kg to 15-17 kg per container. The fish consumed 8 to 1 1 kg of feed per tank, which corresponds to a feed conversion rate (FCR) of 0.8 to 0.9 kg feed per kg of fish.

The results of the feeding experiments are shown in the table below. Table 6: Weight gain of the fish as a function of the diet

Overall, it was found that an increase in the growth of the salmon could be achieved both by complete and partial replacement of the fish oil by a novel Aurantiochytrien- biomass.

Interestingly, when the fish oil was partially replaced by the aurantiochrome biomass, a higher salmon growth was achieved than with complete replacement by the aurochrome biomass.

It was found that the fish fed with the control formula Diet 1, with a final weight of 331 g average a significantly lower

Final weight had as the fish fed with the diet 1 or 2 recipes were. The fish that were fed with the formula Diet 2 performed best: they achieved a significantly higher final weight of 362 g on average.

Example 10: Fatty acid utilization by the fish

To determine the fatty acid utilization, the lipid detection was carried out using the Bligh & Dryer extraction method and subsequent fatty acid analysis according to AOCS Ce 1 b-89. Both muscle samples and whole salmon samples were analyzed. In this case, the results shown in the following tables were obtained (indicated in each case the amount of fats determined at the start or after the end of the diet in grams, in each case with respect to 100 g total fat).

Table 7: Fatty acid profiles of salmon muscle samples as a function of the diet

Table 8: Fatty acid profiles of whole salmon samples as a function of the diet

It can be observed that even with partial replacement of the fish oil by a

According to the invention Aurantiochytrien biomass a significant increase in the content of PUFAs, omega-3 fatty acids and DHA could be achieved. With complete replacement of the fish oil by the aurantiochrome biomass, the increase in the content of PUFAs is correspondingly higher. Example 11: Determination of the fat content of salmon liver

In each case 3 young salmon were fed for 9 weeks each with the different formulations Diet 1, 2 and 3 and then taken to determine the fat content of the liver of the salmon. The fat extraction was carried out according to the method of Folch (1957, J. Biol. Chem., 226 (1), 497-509). The fat content was then determined by a gravimetric method.

It turned out that due to the presence of the biomass according to the invention compared to feeding without biomass, the fat content in the liver of 8% by weight could be significantly reduced to 4-5% by weight.

The accumulation of fat in the liver is considered as an indication of an imbalance of the food metabolism and in particular as an indication of oxidative stress. The significant reduction of fat in the liver is therefore a clear indication of the reduction of stress and thus of the health status of salmon.

Claims

claims

1 . Process for the preparation of biomass containing PUFAs, characterized

characterized in that for the production of the biomass, a cultivation of

Microorganisms are carried out in a fermentation medium containing sulfate in an amount such that in the resulting biomass, a sulfate content, based on the dry mass, of 25 to 60 g / kg, wherein the

Microorganisms around cells of the taxon Labyrinthulomycetes acts.

2. Method according to the preceding claim, characterized in that the fermentation medium contains sulfate in an amount so that in the

resulting biomass a sulfate content, based on the dry matter, from 25 to 50, preferably 25 to 40, particularly preferably 25 to 35 g / kg adjusted.

3. The method according to any one of the preceding claims, characterized in that it is in the microorganisms of the taxon Labyrinthulomycetes

(Labyrinthulea, slime - fungi, mucus) around those of the

Thraustochytriaceae, preferably of the genera Althomia, Aplanochytrium, Elnia, Japonochytrium, Schizochytrium, Thraustochytrium, Aurantiochytrium,

Oblongichytrium or ulkenia, acts.

4. Method according to the preceding claim, characterized in that the microorganisms are cells of the genus Aurantiochytrium, in particular those of the species Aurantiochytrium limacinum, in particular those of the strain Aurantiochytrium limacinum SR21 (IFO 32693).

5. PUFAs containing biomass, characterized in that it has a sulfate content, based on the dry mass, of 25 to 60 g / kg, wherein the biomass cells of the taxon Labyrinthulomycetes.

6. PUFAs containing biomass according to the preceding claim, characterized

in that it has a sulphate content, based on the dry matter, of 25 to 50, preferably 25 to 40, particularly preferably 25 to 30 g / kg.

7. PUFAs-containing biomass according to any one of the preceding claims, characterized in that it is in the cells of the taxon Labyrinthulomycetes (Labyrinthulea, Netzschleimpilze, mucus) to those of the family

Oblongichytrium or ulkenia, acts.

8. PUFAs containing biomass according to the preceding claim, characterized

in that the microorganisms are cells of the genus Aurantiochytrium, in particular those of the species Aurantiochytrium limacinum, in particular those of the strain Aurantiochytrium limacinum SR21 (IFO 32693).

9. A method for producing a feed, characterized in that a PUFAs containing biomass according to one of the preceding claims mixed with other feed components and then extruded into a feed and optionally the resulting extrudate is then coated with oil.

10. The method according to the preceding claim, characterized in that the composition used in the extrusion has the following properties: a) a total protein content of 33 to 67 wt .-%, preferably 39 to 61

Wt .-%, in particular 44 to 55 wt .-%; b) a total fat content of 5 to 25 wt .-%, preferably 8 to 22 wt .-%, in particular 10 to 20 wt .-%, especially 12 to 18 wt .-%; c) a total starch content of not more than 25% by weight, in particular not more than 20% by weight, preferably from 6 to 17% by weight, particularly preferably from 8 to 14% by weight; d) a biomass content, in particular labyrinthulea biomass, preferably Thraustochytriaceae biomass, of 2 to 24% by weight, preferably 4 to 22% by weight, in particular 9 to 20% by weight, especially 1 1 to 18 wt .-%; e) preferably has a content of polyunsaturated fatty acids (PUFAs) of 0.8 to 8 wt .-%, preferably 1, 2 to 6 wt .-%, in particular 1, 4 to 5 wt .-%, especially 1, 5 bis 4% by weight; f) preferably a content of omega-3 fatty acids of 0.8 to 8 wt .-%,

preferably 1, 2 to 6 wt .-%, in particular 1, 4 to 5 wt .-%, especially 1, 5 to 4 wt .-%; g) preferably content of DHA from 0.1 to 4.0 wt .-%, preferably 0.25 to 3.0 wt .-%, in particular 0.5 to 2.8 wt .-%, especially 0.8 to 2.5 wt .-%, in particular 1, 0 to 2.0 wt .-%.

1 1. Method according to one of the preceding claims, characterized in that the extrusion at an energy input of 12 - 28 Wh / kg, in particular 14 - 26 Wh / kg, particularly preferably 16 - 24 Wh / kg, is performed.

12. The method according to any one of the preceding claims, characterized in that after the extrusion and optionally drying of the extrudate

Coating of the extrudate with oil, in particular vegetable oil, preferably in an amount of, with respect to the final product, 3 to 17 wt .-%, in particular 5 to 15 wt .-%, takes place.

13. feed, characterized in that it contains a PUFAs containing biomass according to one of claims 1 to 8.

14. Feed according to claim 13, characterized in that it has the following

Properties has: a) a total protein content of 30 to 60 wt .-%, preferably 35 to 55 wt .-%, in particular 40 to 50 wt .-%; b) a total fat content of 15 to 35 wt .-%, preferably 18 to 32

Wt .-%, in particular 20 to 30 wt .-%, especially 22 to 28 wt .-%; c) a total starch content of not more than 25% by weight, in particular not more than 20% by weight, preferably from 5 to 15% by weight, more preferably from 7 to 13% by weight; d) a biomass content, in particular of labyrinthulea biomass,

preferably to Thraustochytriaceae biomass, from 2 to 22 wt .-%, preferably 4 to 20 wt .-%, in particular 8 to 18 wt .-%, especially 10 to 16 wt .-%; e) preferably a content of polyunsaturated fatty acids (PUFAs) of 2 to 12 wt .-%, preferably 3 to 10 wt .-%, in particular 4 to 9 wt .-%, especially 5 to 8 wt .-%; f) preferably has a content of omega-3 fatty acids of 1 to 6 wt .-%, preferably 1, 5 to 5 wt .-%, in particular 2 to 4.5 wt .-%, especially 2.5 to 4 wt .-%; g) preferably has a content of DHA of 0.5 to 3 wt .-%, preferably 0.8 to 2.5 wt .-%, in particular 1 to 2.5 wt .-%, especially 1, 2 to 2, 2 parts by weight

%, in particular 1, 2 to 2.0 wt .-%.

15. A method for growing animals, especially fish, in which a

Feed according to one of the preceding claims is used.